The objective of this project is a thorough investigation of the functional and structural aspects of hepatic subcellular organelles after acute and chronic ethanol intoxication. This is supplemented by measuring the effects of ethanol, and its primary metabolite, acetaldehyde, in vitro. Together with Dr. C. S. Lieber, we produced alcoholic cirrhosis in baboons within two years, and we are now engaged in studies designed to quantitate the transition of fatty liver to cirrhosis. We have shown that certain drugs which affect alcohol metabolism, namely, pyrazol and clofibrate, have potent effects upon mitochondria, including inhibition of fatty acid oxidation and the shuttles for the transport of reducing equivalents. The persistent effect of chronic ethanol intoxication on mitochondrial electron transport seems to involve complex 1 of the respiratory-phosphorylation chain. During the course of our studies we have developed new techniques for isolating cell organelles, including cation-bound microsomes, which sediment rapidly at 30Xg, and a method for isolating plasma membranes in the same preparation used for mitochondria and microsomes. We will continue the current studies, stressing subcellular organelles in baboons fed ethanol, and the use of isolated hepatic parenchymal cells.